The Parasitological Institute of the Slovak Academy of Sciences and the Section of Parasitology (formerly Institute of Parasitology) of the Department of Public Health Sciences of the Sapienza University of Rome have had a long term collaboration within several projects, starting with the first visiting fellowships in 1992 in Rome. Since then, the collaboration between the two institutions has continuously developed and has been focused mainly on the systematics of cestodes, starting in particular with the studies on the genetic polymorphism of fish Proteocephalus tapeworms in different host populations examined by enzyme electrophoresis. Some molecular studies have been later carried out on Eubothrium spp. and Echinococcus multilocularis, and most of our joint interest has been focused on Echinococcus granulosus and more recently, on Ascaris nematodes. The major achievements obtained during these years of collaboration are herein reported.
V. Šnábel, M. Miterpáková, S. D’Amelio, M. Busi, D. Bartková, L’. Turčeková, C. Maddox-Hyttel, P. Skuce and P. Dubinský
Nucleotide sequencing of the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene and isoenzyme analysis were used to survey the genetic variability in Echinococcus multilocularis populations from Slovakia. A sample of 12 isolates acquired from 10 different districts from red foxes exhibited identical sequences. Compared with the previously described E. multilocularis variants, one base substitution was consistently observed relative to the M1 variant (detected in China, Alaska, North America, Japan) and three base substitutions were recorded relative to the M2 variant (detected in Germany) in the CO1 fragment. These data, along with the recently gathered data from French isolates, are indicative of a genetically unique population occurring in Central and Western Europe. Electrophoretic examination of enzymes produced by 14 gene loci revealed intraspecific polymorphism only with the glucose-phosphate isomerase (two distinct patterns) and the mannosephosphate isomerase (four genotypes composed of three alleles) enzyme systems. To allow a fast species differentiation of E. multilocularis and E. granulosus (specifically, the G7 genotype occurring in Slovakia), discriminative electrophoretic characters between the species were obtained by isoenzyme analysis. Fixed genetic differences between the species were detected in the glucose-phosphate isomerase, esterase and aldolase systems, and partial differences were detected in four additional systems.
S. Cavallero, R. A. El Sherif, A. Pizzarelli, A.A. El Fituri, M. El Showhdi, F. Benmosa and S. D’Amelio
The occurrence of zoonotic parasitic nematodes in Atlantic chub mackerels (Scomber colias syn. Scomber japonicus) from Libyan waters was investigated, using epizootiological estimations and molecular specific characterization of larvae. Nematodes belonging to Anisakis spp., the main etiological agent of anisakiasis in Mediterranean waters, and to Hysterothylacium spp. so far considered not pathogenic to humans, were detected. Prevalence values were generally high in visceral cavities (over 40 % for both parasites) while were low for Anisakis (around 1 %) and null for Hysterothylacium in muscles. Moreover, the level of infections was associated with seasons, a feature potentially useful to plan fishing captures and to elaborate risk mitigation strategies for anisakiasis. Species molecular identification performed on a subsample described the presence of Hysterothylacium aduncum as the predominant species, along with Anisakis pegreffii and the hybrids (A. pegreffii and A. simplex sensu stricto), thus posing a concrete zoonotic risk following the consumption of such fish species as a raw preparation.
V. Šnábel, C. Calma, S. O. Georgescu, S. Cavallero, S. D’Amelio, Z. Vasilková and A. Ito
Cystic echinococcosis (CE) and alveolar echinococcosis (AE) are considered as one of the most important zoonotic diseases in Romania, where they are subject to mandatory reporting. To obtain more knowledge about the genetic diversity of Echinococcus causative agents of these diseases, 11 isolates from humans and ungulate intermediate hosts from the two regions of Romania were genotyped using mitochondrial markers. In clinical samples of five patients from north-eastern Romania (Iasi, Botosani, Vaslui counties), Echinococcus multilocularis was identified as causal agent by cox1 sequence analysis. To the best of our knowledge this finding presents the first molecular evidence of E. multilocularis in humans from Romania. Only two cases of AE in patients were previously documented in the country by serological methods. In our four patients the most widespread European variant E5 of E. multilocularis was recorded, whereas in isolate from Vaslui county three nucleotide substitutions were detected as compared to the most related E5 haplotype. One of these mutations (411T/G) matched N1 and N2 haplotypes described previously from North America. In six CE samples retrieved from western Romania (Caras-Severin and Timis counties), two human isolates were diagnosed as Echinococcus canadensis G7, one as E. granulosus s.s. G1 and one as E. granulosus s.s. G3 using atp6 and rrnS sequencing. In ungulates, the cattle isolate was allocated to E. granulosus s.s. G1 and pig isolate to E. canadensis G7. The two G7 findings in humans reinforced the recent view that G7 was underestimated as compared to the E. granulosus s.s. regarding human CE threat that can be further employed for identifying sources of infections and establishing suitable preventive measures.